An Ammonia-Induced Calcium Phosphate Nanostructure: A Potential Assay for Studying Osteoporosis and Bone Metastasis

ACS Appl Mater Interfaces. 2021 Apr 21;13(15):17207-17219. doi: 10.1021/acsami.1c00495. Epub 2021 Apr 12.

Abstract

Osteoclastic resorption of bones plays a central role in both osteoporosis and bone metastasis. A reliable in vitro assay that simulates osteoclastic resorption in vivo would significantly speed up the process of developing effective therapeutic solutions for those diseases. Here, we reported the development of a novel and robust nanostructured calcium phosphate coating with unique functions on the track-etched porous membrane by using an ammonia-induced mineralization (AiM) technique. The calcium phosphate coating uniformly covers one side of the PET membrane, enabling testing for osteoclastic resorption. The track-etched pores in the PET membrane allow calcium phosphate mineral pins to grow inside, which, on the one hand, enhances coating integration with a membrane substrate and, on the other hand, provides diffusion channels for delivering drugs from the lower chamber of a double-chamber cell culture system. The applications of the processed calcium phosphate coating were first demonstrated as a drug screening device by using alendronate, a widely used drug for osteoporosis. It was confirmed that the delivery of alendronate significantly decreased both the number of monocyte-differentiated osteoclasts and coating resorption. To demonstrate the application in studying bone metastasis, we delivered a PC3 prostate cancer-conditioned medium and confirmed that both the differentiation of monocytes into osteoclasts and the osteoclastic resorption of the calcium phosphate coating were significantly enhanced. This novel assay thus provides a new platform for studying osteoclastic activities and assessing drug efficacy in vitro.

Keywords: ammonia-induced calcium phosphate insert; drug delivery; osteoclasts; prostate cancer; resorption assay.

MeSH terms

  • Alendronate / administration & dosage
  • Alendronate / pharmacology
  • Ammonia / chemistry*
  • Animals
  • Bone and Bones / drug effects
  • Bone and Bones / pathology*
  • Calcium Phosphates / chemistry*
  • Cell Differentiation / drug effects
  • Drug Carriers / chemistry
  • Humans
  • Membranes, Artificial
  • Mice
  • Nanostructures / chemistry*
  • Osteoclasts / drug effects
  • Osteoclasts / pathology
  • Osteoporosis / metabolism
  • Osteoporosis / pathology
  • Osteoporosis / physiopathology*
  • PC-3 Cells
  • Porosity
  • RAW 264.7 Cells

Substances

  • Calcium Phosphates
  • Drug Carriers
  • Membranes, Artificial
  • Ammonia
  • calcium phosphate
  • Alendronate